Müller Patricia, Seyfried Patrick, Frühauf Anton, Heckel Alexander
Institute for Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt am Main, Germany.
Institute for Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt am Main, Germany.
Methods Enzymol. 2019;624:89-111. doi: 10.1016/bs.mie.2019.04.019. Epub 2019 May 7.
The ability to address the function of oligonucleotides with light is highly desirable since they are often used experimentally in the regulation of biological processes that need to be controlled in time, space and activation level. Here we present an extension of our initial approach of using photo-tethers that force single strands of nucleic acids into a circle, thus making them unable to form a duplex with a complementary DNA- or RNA-strand. Due to the persistence length a single strand can form a circle of, for example, 30 nucleotides, but a duplex cannot. We show that these new photo-tethers can also be easily installed on the phosphodiester backbone. This simplifies the approach considerably and leads to temporarily inhibited oligonucleotides that can only form a duplex after linearization by photoactivation.
利用光来调控寡核苷酸功能的能力非常令人期待，因为寡核苷酸常用于实验中对生物过程的调控，而这些生物过程需要在时间、空间和激活水平上进行控制。在此，我们展示了对我们最初方法的扩展，即使用光连接子迫使单链核酸形成环状，从而使其无法与互补的DNA或RNA链形成双链体。由于持久长度，单链可以形成例如30个核苷酸的环，但双链体则不能。我们表明，这些新的光连接子也可以很容易地安装在磷酸二酯主链上。这极大地简化了该方法，并产生了只能在光激活使其线性化后才能形成双链体的暂时受抑制的寡核苷酸。
